To monitor and analyze Cricetulus migratorius fungal diversity ,60 adult Cricetulus migratorius brought from Xinjiang region of China were dissected after being euthanized and the specimens were collected .Fungal diversity research was carried out by TaqMan MGB probe real-time fluorescence quantitative PCR ,the ribosome cloning sequencing ,and fungal cul-ture identification techniques .The 60 fungal isolates were characterized from Cricetulus migratorius ,including Candida albi-cans ,Trichosporonasahii ,Aspergillus fumigatus ,Aspergillusniger ,Aspergillussydowii ,Aspergillus japonicus ,Asper-gillus ustus ,Aspergillus versicolor ,Penicillium chrysogenum ,Paecilomyces variotii ,Penicillium aurantiogriseum ,Neuros-pora sitophila ,Neurospora intermedia ,and Cladosporium cladosporioides .Many of them associated with grey hamster as zoonotic pathogens .The results showed that the most dominant fungal group was Aspergillus ,and Penicillium followed by it . These fungi were susceptible to nystatin ,clotrimazole and voriconazole .It’s indicated that application of TaqMan MGB probe real-time fluorescence quantitative PCR ,the ribosome cloning sequencing ,and fungal culture identification techniques could ef-fectively analyze Cricetulus migratorius .The results could provide a scientific basis for Cricetulus migratorius microbiological monitoring and quality standard establishment in China .Overall ,the findings of the present study constitute ,to the authors’ knowledge ,the first extensive report on the diversity of fungal flora associated with Cricetulus migratorius .

Objective Giardia lamblia is an important pathogen of zoonosis giardiasis , it poses a potential threat to the quality of SPF (specific pathogen-free) laboratory animals cannot be ignored.The aim of this study is to establish the method of rapid diagnosis of Giardia lamblia, and analyze the test results of 516 batches form 17 manufactures.Methods Direct microscopy, Giemsa-fast staining and multiplex polymerase chain reaction (multiplex PCR) were applied to detect Giardia lamblia.Results Numerous of Giardia lamblia trophozoites and cysts were detected in SPF laboratory animals by using direct microscopy and Giemsa-fast staining, and multiplex PCR were performed to identify 18S rDNA,β-giardin, TPI and GDH genes of DNA extracted from these trophozoites and cysts identified Giardia lamblia.Direct microscopy, Giemsa-fast staining, and multiplex PCR methods can be used to detect Giardia lamblia.Of the 2562 SPF laboratory animals studied, 22.9%(586/2562) were positive for Giardia lamblia.Conclusions Direct microscopy , Giemsa-fast staining , and multiplex PCR were effective techniques with high sensitivity and specificity for rapid diagnosis of Giardia lambliain.It is not satisfactory that the results of Giardia lamblia examination in 516 batches form 17 manufactures failed to meet the requirements 100%.

Objective To diagnosis tumor transplanted nude mice Strongyloidiasis .Methods Postmortem microscopic examination of the tumor transplanted nude mice detected Strongyloides stercoralis for morphological identification and double polymerase chain reaction ( PCR ) assay for molecular diagnosis of Strongyloides stercoralis infection in tumor transplanted nude mice .Results Presence of numerous S .stercoralis in autopsy in tumor transplanted nude mice samples preliminary determined the diagnosis of strongyloidiasis .Confirmed diagnosis of Strongyloides stercoralis infection by double PCR detection of specific DNA in tumor transplanted nude mice samples .Conclusion The most important clue to prevent such serious consequences is early diagnosis .Tumor transplanted recipients and donors should be screened for parasitic infections including strongyloidiasis .To the authors ’ knowledge , this study is the first extensive report on diagnosis tumor transplanted nude mice Strongyloidiasis .

To investigate the prevalence and the antibiotic resistance of bacteria isolated from 25 miniature pigs. 45 bacterial strains were isolated, which were identified by biochemical assays, amplification of 16S rRNA genes by PCR and sequence analysis, and were evaluated for resistance to 30 antibiotics. The identification results showed that these bacteria belonged to Campylobacter (Campylobacter jejuni), Helicobacterium (Helicobacter pylori), Klebsiella (Klebsiella pneumoniae), Escherichia (Escherichia coli, Escherichia fergusonii), Pseudomonas (Pseudomonas aeruginosa), Stenotrophomonas (Stenotrophomonas maltophilia), Staphylococcus (Staphylococcus aureus, Staphylococcus haemolyticus, Staphylococcus simulans), Streptococcus (Streptococcus pneumoniae, Streptococcus suis, Streptococcus vestibularis, Streptococcus mitis, Gemella measles, Aerococcus viridans) and Bacillus (Bacillus subtilis, Bacillus licheniformis, Bacillus alvei, Bacterium megaterium). These bacteria were all susceptible to aztreonam and cephalothin. However, the resistence to furazolidone was found. Microbial population carried by miniature pigs in China had characters of diversity. Results of this study provided scientifical accordance for the microorganism monitoring of miniature pigs in China.

Objective To evaluate the therapeutic effects of methamphetamine (MA) dependence and the repairment of DA neuronal function by SPECT corpus striatum DAT visual digital neural molecular imaging techniques.Methods 25 MA dependent patients (BPRS score ≥ 35) were treated by self-designed treatment program for more than 6 months.The clinical therapeutic effects were scored with reducing rate of BPRS.MA dependent patients were examined by SPECT corpus striatum DAT imaging before and after treatment,while healthy volunteers were examined only once.The SPECT corpus striatum DAT images were analyzed visually and quantitatively.Results The reducing rate of BPRS showed that the total effective rate was 80.0％.Visual analysis of SPECT corpus striatum DAT images showed that the distribution of DAT in the corpus striatum was regionally reduced or defected in various degrees before treatment,and was significantly increased after treatment.Quantitative analysis showed that the bilateral striatal V ((19.26 ± 2.85) cm3),m((20.22±2.99) g) and Ra(4.78±0.79) ％) of MA dependent patients were significantly lower compared with those of the healthy volunteers(respectively (35.39±4.42) cm3,(37.16±4.64) g and (7.93± 0.86) ％) (all P＜ 0.01) before treatment and were significantly improved (P＜ 0.01) after treatment (V:(22.80±4.28) cm3,m:(23.93± 4.49) g and Ra:(5.64 ± 0.99) ％) with a 76.0％ corpus striatum DAT improvement rate.However,the bilateral striatal V,m and Ra of MA dependent patients after treatment were still lower than those of the healthy volunteers (P＜0.01).There was no significant difference between the striatal DAT improvement rate and the BPRS reduction rate (P＞ 0.05).Conclusion SPECT corpus striatum DAT visual digital neural molecular imaging techniques are reliable in the evaluation of the treatment programs for MA dependence and the repair of DA neuronal function.

Objective To investigate the electrophysiological properties of neurons in visual cortex of binocular form-deprivated rats during the developmental stages and to explore the synaptic and cellular mechanisms of plasticity in the visual cortex. Methods Whole cell patch clamp technique was applied to the visual cortex slices prepared from 61 Wistar rats of both genders, which were divided into two groups; control group and binocular form-deprivation (BFD) group. Excitatory postsynaptic currents (EPSCs) and long term potentiation (LTP) of the neurons were recorded. Results (1) The percentage of silent response in BFD group was significantly higher than that in the control group(P

Objective According to the basic theory of Yin and Yang,acute urticaria should be treated with divergent medicine instead of convergence medicine.In order to prove the theory,we carry out the quantifiable and repeatability traditional Chinese medicine screening tests in acute urticaria patients,with double controls of experimental group (divergent Chinese medicine),control group (convergence Chinese medicine)end instrumental quality-control standard substance MEBE.Methods Germany MORA-Super bio-resonance instrument (medical mode) as adopted as objective index.32 cases of acute phase urticaria were selected and treated with divergent or convergence Chinese traditional medicine,to observe the results whether conform to traditional Chinese medicine basic theory of Yin and Yang (That is whether the acute phase urticaria should be treated with divergent Chinese medicine).Results Through the 32 cases repeat experiments,22 out of 23 kinds of Divergent Chinese medicine have been screened out in experimental group; while none has been screened out in 12 kinds of convergence Chinese medicine in control group.There was significant difference between two groups (P＜0.01).The result was entirely consistent with TCM basic theory of opposite relationship between yin and yang.The top five divergent Chinese medicines were divaricate saposhnikovia root,cicada slough,catnip,shrub chastetree rruit,and lily magnolia.Conclusion Treated acute urticaria with divergent Chinese medicine conforms to the basic theory of traditional Chinese medicine.

Objective To study the immunoreaction of the recombinant proteins encoded by the fragments of ORF2 ( second open reading frames) gene of hepatitis E virus ( HEV). Methods The aimed sequence from the full-length ORF2 in clone PEH2 , which was derived from a Chinese strain of HEV,was amplified and cloned it into vector pcDNAS. 1 which was then transfected to 293 cell. Results The ORF2 protein was present in the soluble fractions of the cell lysate. The expressed protein of HEV ORF2 in 293 cells by using a plasmid pcDNA3. 1-based system showed positive on immunoblots probed against antibodies raised in BALB/c mice. Conclusion The experimental results laid a foundation for developing diagnostic reagent to detect HEV by using the expressed products of ORF2 protein.

Objective To develop a TaqMan MGB probe-based,sensitive and specific real-time fluorescence quantitative PCR assay for rapid detection of Helicobacter hepaticus.Methods Primers and probes specific toflaB gene of Helicobacter hepaticus were designed.A TaqMan MGB probe-based,real-time fluorescence quantitative PCR was established.The specificity,sensitivity and stability of the assay were assassed.Then,the established TaqMan MGB probe real-time fluorescence quantitative PCR assay was applied to detect Helicobacter hepaticus in 1081 clinical specimens during 2008-2011,compared with bacterial isolation and culture method and conventional PCR assay.Results The specificity of this established TaqMan MGB probe-based real-time fluorescence quantitative PCR was high and there were no cross-reactivity with Helicobacter pylori,Campylobacter jejuni,Clostridium piliforme,Pasteurella pneumotropica,Escherichia coli,Pseudomonas aeruginosa.The detection limits was 8.3 copies.The correlation coefficient and slope value of standard curve were 0.999 and -3.227,respectively and the efficiency of TaqMan MGB-based probe realtime fluorescence quantitative PCR assay was 100％.The TaqMan MGB-based probe real-time fluorescence quantitative PCR and conventional PCR were preformed to detect Helicobacter hepaticus in 1081 clinical specimens,a total of 86 specimens were positive for Helicobacter hepaticus.However,there was only 4 specimens were positive by bacteria isolation and culture method.The results showed that TaqMan MGB -based probe real-time fluorescence quantitative PCR for Helicobacter hepaticas was more sensitive than bacteria isolation and culture method,and it could detect Helicobacter hepaticus DNA from clinical specimens directly,and detection time is only 2 hours.Conclusion The TaqMan MGB-based probe real-time fluorescence quantitative PCR assay was a reliable,specific,sensitive and useful tool for rapid detection of Helicobacter hepaticus.

Objective To analyze and evaluate the population genetic quality of 3 subbreeds of China Agricultural University miniature pigs in Beijing.Method According to the local standard DB11/T828.3 -2011, 25 pairs of microsatellite primers were used in 3 subbreeds of China Agricultural University miniature pigs, and software Popgen32 was used to process the data.Results 24 microsatellite loci shared 130, 122 and 138 alleles in the China Agricultural University miniature pigs I, II, III, respectively. The average heterozygosity was 0.6759, 0.5967 and 0.6779, respectively, while the average polymorphism information content ( PIC) was 0.6344, 0.5540 and 0.6403, respectively. The genetic distance between China Agricultural University miniature pig II and III was 0.4251, while the genetic distance between China Agricultural University miniature pig I and II was 0.2084.Conclusions In the 3 subbreeds, China Agricultural University miniature pigs II and III have genetic stability and genetic diversity, and both of which satisfy with the genetic characteristics of closed colony laboratory animal.